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Nitrosylation precedes caspase-3 activation and translocation of apoptosis-inducing factor in neonatal rat cerebral hypoxia-ischaemia

Artikel i vetenskaplig tidskrift
Författare Changlian Zhu
Xiaoyang Wang
L. Qiu
C. Peeters-Scholte
Henrik Hagberg
Klas Blomgren
Publicerad i J Neurochem
Volym 90
Nummer/häfte 2
Sidor 462-71
Publiceringsår 2004
Publicerad vid Institutionen för kvinnors och barns hälsa, Avdelningen för obstetrik och gynekologi
Institutionen för kvinnors och barns hälsa, Avdelningen för pediatrik
Institutionen för fysiologi och farmakologi, Avdelningen för fysiologi
Sidor 462-71
Språk en
Länkar www.ncbi.nlm.nih.gov/entrez/query.f...
Ämnesord Animals, Animals, Newborn, Apoptosis Inducing Factor, Biological Markers/analysis, Biotin/*analogs & derivatives/pharmacology, Caspases/*metabolism, Cell Count, DNA Damage, Disease Models, Animal, Enzyme Activation/drug effects, Enzyme Inhibitors/pharmacology, Female, Flavoproteins/*metabolism, Hypoxia-Ischemia, Brain/enzymology/*metabolism/pathology, Male, Membrane Proteins/*metabolism, Neurons/drug effects/metabolism/pathology, Nitric Oxide Synthase/antagonists & inhibitors, Nitric Oxide Synthase Type I, Nitric Oxide Synthase Type II, Protein Transport/physiology, Rats, Rats, Wistar, Research Support, Non-U.S. Gov't, Tyrosine/*analogs & derivatives/analysis/biosynthesis/*metabolism
Ämneskategorier Experimentell hjärnforskning, Neurobiologi

Sammanfattning

Excessive nitric oxide (NO) production after cerebral hypoxia-ischaemia (HI) may induce cellular injury in various ways, including reaction with superoxide to form the highly reactive peroxynitrite. We characterized the spatial and temporal formation of peroxynitrite through immunohistochemical detection of nitrosylated proteins. Nitrotyrosine immunoreactivity peaked around 3 h post-HI and was detected in areas of injury, as judged by the loss of microtubule-associated protein-2 (MAP-2) staining, in neurones, glia and endothelial cells. Nitrotyrosine staining co-localized with three other cellular markers of injury, active caspase-3, nuclear translocation of apoptosis-inducing factor (AIF) and an oligonucleotide hairpin probe detecting specific DNA strand breaks. The number of nitrotyrosine-positive cells at early time points outnumbered the cells positive for the other three markers of injury, indicating that nitrosylation preceded caspase-3 activation. Pharmacological inhibition of neuronal and inducible nitric oxide synthase (nNOS and iNOS) using 2-iminobiotin, which has been demonstrated earlier to be neuroprotective, significantly reduced nitrotyrosine formation and caspase-3 activation, but not nuclear translocation of AIF, in cortex and striatum of the ipsilatral hemisphere. In summary, nitrotyrosine is an early marker of cellular injury and inhibition of nNOS and iNOS is a promising strategy for neuroprotection after perinatal HI.

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